Search Results (10)

Cryopreservation is a critical strategy for the long-term conservation of plant germplasm, particularly for clonally propagated crops, endangered species, and plants producing recalcitrant seeds. Hydrogel-based encapsulation systems can improve survival during ultra-low-temperature storage by providing mechanical protection, moderating dehydration, and regulating cryoprotectant uptake. Although calcium–alginate beads remain the traditional matrix for encapsulation–dehydration and encapsulation–vitrification, recent advances in biomaterials science have enabled the development of composite polysaccharide blends, protein-based matrices, synthetic polymer networks, macroporous cryogels, and functionalized hybrid hydrogels incorporating surfactants, antioxidants, or nanomaterials. These engineered systems provide improved control over water state, pore architecture, diffusion kinetics, and thermal behavior, thereby reducing cryoinjury and enhancing post-thaw recovery across diverse plant explants. This review synthesizes current knowledge on hydrogel platforms used in plant cryopreservation, with emphasis on how physicochemical properties influence dehydration dynamics, cryoprotectant transport, vitrification stability, and rewarming responses. Performance across major explant types is assessed, key limitations in existing materials and protocols are identified, and design principles for next-generation hydrogel systems are outlined. Future progress will depend on material standardization, integration with automated cryopreservation workflows, and the development of responsive hydrogel matrices capable of mitigating cryogenic stresses. Full article

Rhodomyrtus psidioides (G.Don) Benth. (Myrtaceae) is a critically endangered rainforest species from the east coast of Australia, where populations have severely and rapidly declined due to the effects of repeated myrtle rust infection. With very limited material available in the wild and freezing-sensitive seeds that have prevented storage in a seed bank, ex situ conservation of this exceptional species has proven difficult. Material from a seed orchard grown at the Australian Botanic Garden Mount Annan was successfully used to initiate three new accessions into tissue culture from cuttings, and to undertake cryopreservation experiments using a droplet-vitrification (DV) protocol for both seeds and cultured shoot tips. Use of seedling material for tissue culture initiation was very effective, with a 94–100% success rate for semi-hardwood explants and a 50–62% success rate for softwood explants. Although no survival of seeds after cryopreservation was observed, seeds of R. psidioides showed some tolerance of desiccation and exposure to cryoprotective agents. Regeneration after cryopreservation using a DV protocol was demonstrated in only one shoot tip precultured on basal medium containing 0.4 M sucrose and incubated in PVS2 for 20 min prior to immersion in liquid nitrogen. These results demonstrate the value of living collections in botanic gardens for conservation research, highlight the importance of germplasm choice for tissue culture initiation, and demonstrate the potential of cryobiotechnologies for the ex situ conservation of exceptional plant species. Full article

The lifespan or longevity of a seed is the time period over which it can remain viable. Seed longevity is a complex trait and varies greatly between species and even seed lots of the same species. Our scientific understanding of seed longevity has advanced from anecdotal ‘Thumb Rules,’ to empirically based models, biophysical explanations for why those models sometimes work or fail, and to the profound realisation that seeds are the model of the underexplored realm of biology when water is so limited that the cytoplasm solidifies. The environmental variables of moisture and temperature are essential factors that define survival or death, as well as the timescale to measure lifespan. There is an increasing understanding of how these factors induce cytoplasmic solidification and affect glassy properties. Cytoplasmic solidification slows down, but does not stop, the chemical reactions involved in ageing. Continued degradation of proteins, lipids and nucleic acids damage cell constituents and reduce the seed’s metabolic capacity, eventually impairing the ability to germinate. This review captures the evolution of knowledge on seed longevity over the past five decades in relation to seed ageing mechanisms, technology development, including tools to predict seed storage behaviour and non-invasive techniques for seed longevity assessment. It is concluded that seed storage biology is a complex science covering seed physiology, biophysics, biochemistry and multi-omic technologies, and simultaneous knowledge advancement in these areas is necessary to improve seed storage efficacy for crops and wild species biodiversity conservation. Full article

Holm oak populations are severely affected by oak decline syndrome, and reliable methods of conserving the plant material are required. A vitrification-based cryopreservation method was used for the first time for the long-term conservation of holm oak embryogenic cultures. Successful cryopreservation was achieved after determining the best developmental stage of the somatic embryos used and the optimal incubation period in plant vitrification solution 2 (PVS2). Embryos were recovered from individual nodular embryogenic structures (NES) derived from four embryogenic lines after preculture on a medium containing 0.3 M sucrose, incubation in PVS2 vitrification solution for 15 min at 25 °C and direct immersion in liquid nitrogen (LN). Embryo recovery rates of 16.7–63.3% were obtained after cryostorage for four years in LN. In addition to the embryo developmental stage and the PVS2 treatment time, the genotype can also significantly affect embryo recovery after LN storage. There were no significant differences in plant regeneration or polyploid stability between somatic embryos and plants derived from control embryos (not cryopreserved) and cryopreserved embryos. The findings indicate that embryo proliferation, plant conversion and polyploid stability are maintained in material recovered from the vitrification solution and subsequently cryopreserved. Full article

The Myrtaceae is a very large and diverse family containing a number of economically and ecologically valuable species. In Australia, the family contains approximately 1700 species from 70 genera and is structurally and floristically dominant in many diverse ecosystems. In addition to threats from habitat fragmentation and increasing rates of natural disasters, infection by myrtle rust caused by Austropuccinia psidii is of significant concern to Australian Myrtaceae species. Repeated infections of new growth have caused host death and suppressed host populations by preventing seed set. Although most Myrtaceae species demonstrate orthodox seed storage behavior, exceptional species such as those with desiccation sensitive seed or from myrtle rust-suppressed populations require alternate conservation strategies such as those offered by cryobiotechnology. Targeting seven key Australian genera, we reviewed the available literature for examples of cryobiotechnology utilized for conservation of Myrtaceae. While there were only limited examples of successful cryopreservation for a few genera in this family, successful cryopreservation of both shoot tips and embryonic axes suggest that cryobiotechnology provides a viable alternative for the conservation of exceptional species and a potential safe storage method for the many Myrtaceae species under threat from A. psidii. Full article

Plant species conservation through cryopreservation using plant vitrification solutions (PVS) is based in empiricism and the mechanisms that confer cell integrity are not well understood. Using ESI-MS/MS analysis and quantification, we generated 12 comparative lipidomics datasets for membranes of embryogenic cells (ECs) of Magnolia officinalis during cryogenic treatments. Each step of the complex PVS-based cryoprotocol had a profoundly different impact on membrane lipid composition. Loading treatment (osmoprotection) remodeled the cell membrane by lipid turnover, between increased phosphatidic acid (PA) and phosphatidylglycerol (PG) and decreased phosphatidylcholine (PC) and phosphatidylethanolamine (PE). The PA increase likely serves as an intermediate for adjustments in lipid metabolism to desiccation stress. Following PVS treatment, lipid levels increased, including PC and PE, and this effectively counteracted the potential for massive loss of lipid species when cryopreservation was implemented in the absence of cryoprotection. The present detailed cryobiotechnology findings suggest that the remodeling of membrane lipids and attenuation of lipid degradation are critical for the successful use of PVS. As lipid metabolism and composition varies with species, these new insights provide a framework for technology development for the preservation of other species at increasing risk of extinction. Full article

Conservation of threatened flora in genetic collections ex situ using in vitro culture and cryopreservation is receiving an increasing recognition as a complementary strategy to in situ conservation in natural habitats. The present study is focused on an integrated approach which involves conservation and propagation, emphasizing the usefulness of cryopreservation techniques for germplasm conservation of streambank lupine (Lupinus rivularis Douglas ex Lindl.), an endangered species in Canada. This included in vitro seed germination on Murashige and Skoog basal medium supplemented with 1 µM thidiazuron to induce multiple shoot formation, micropropagation on a medium with 5 µM benzylaminopurine, and in vitro rooting on medium with 20.0 µM indole-3-butyric acid. Cryopreservation of seeds and shoot tips of in vitro grown plants was successful with over 60% seed germination and 62% regrowth of cryopreserved shoot tips, respectively. Plants developed from cryopreserved seeds had chlorophyll contents in leaves and the growth characteristics including the development of inflorescence, similar to plants raised from non-cryopreserved seeds. These results provide further evidence that the combination of micropropagation with cryopreservation of seeds and vegetative parts may effectively facilitate long-term preservation of L. rivularis and other endangered species. Full article

Conventional dry seed storage is unlikely for about one third of all tree species (and nearly half of evergreen rain forest trees) as they probably produce desiccation sensitive (recalcitrant) seeds. Consequently, international ex situ conservation targets for threatened trees will be difficult to achieve without innovation, especially in cryobiotechnology. We assessed progress in the development of various cryobiotechnology approaches for the preservation of oaks (Quercus), which are keystone species of functioning landscapes, important to the bioeconomy and under increasing threats from the spread of pests and diseases under a changing climate. Various tissues of oaks can be used for banking, from pollen grains to embryo axes. Pollen from five oak species have been shown to be highly desiccation tolerant, making dry pollen storage at low temperatures (including in liquid nitrogen) a valuable technology to support conservation and breeding programs. Somatic embryo (SE) technology and/or shoot tip in vitro technology is available for 39 species, and SE cryopreservation is routinely performed on three commercial species and shoot tips cryopreservation successful in two more species. Seed embryonic axes are the preferred explants for oak ex situ conservation, with tissue survival and regeneration of plants after cryopreservation recorded for 14 and seven species respectively; although differential responses between the shoot and root meristems in the axes are known. Dormant bud preservation seems promising, but is under-researched. Overall, these results indicate the possibility of establishing an integrated platform for the ex situ conservation of oak species based on cryobiotechnology. Challenges of explant choice, optimization of methodologies and large-scale application do remain. However, multiple approaches for the cryopreservation of oak genetic resources are available and implementation programmes should not be delayed, particularly in the centres of species diversity. Full article

The conservation of crop genetic resources, including their wild relatives, is of utmost importance for the future of mankind. Most crops produce orthodox seeds and can, therefore, be stored in seed genebanks. However, this is not an option for crops and species that produce recalcitrant (non-storable) seeds such as cacao, coffee and avocado, for crops that do not produce seeds at all; therefore, they are inevitably vegetatively propagated such as bananas, or crops that are predominantly clonally propagated as their seeds are not true to type, such as potato, cassava and many fruit trees. Field, in vitro and cryopreserved collections provide an alternative in such cases. In this paper, an overview is given on how to manage and setup a field, in vitro and cryopreserved collections, as well as advantages and associated problems taking into account the practical, financial and safety issues in the long-term. In addition, the need for identification of unique accessions and elimination of duplicates is discussed. The different conservation methods are illustrated with practical examples and experiences from national and international genebanks. Finally, the importance of establishing safe and long-term conservation methods and associated backup possibilities is highlighted in the frame of the global COVID-19 pandemic. Full article

Agriculture has always been an important part of human evolution. Traditionally, farming is changing and developing with regard to challenges it faces. The major challenges of modern agriculture are food and nutrition safety for the growing world population. Promoting species and genetic diversity in agriculture appears to be an important approach to dealing with those challenges. Gene banks all around the world play a crucial role in preserving plant genetic resources for future crop improvements. The plant germplasm can be preserved in different ways, depending on the species or form of stored plant tissue. This review focuses on a special preservation method—cryopreservation. Cryopreservation is an effective technique for storing living systems at ultra-low temperatures, usually in liquid nitrogen or its vapor phase. This conservation method is crucial for plants that do not produce seeds or that produce non germinating seeds, as well as for plants that propagate vegetatively. Moreover, based on the cryopreservation method, a novel plant biotechnology tool for pathogen eradication called cryotherapy has been developed. The use of liquid nitrogen eliminates plant pathogens such as viruses, phytoplasmas, and bacteria. Our article reviews recent advances in cryo-biotechnologies such as cryopreservation and cryotherapy, with special focus on studies concerning fruit plants. Full article